Gel Filtration Chromatography Molecular Weight Determination Service

Gel Filtration Chromatography Molecular Weight Determination Service estimates sample molecular weight based on differences in their flow rates through the chromatographic column packed with molecular sieving. Gel filtration chromatography (GFC) offers several unique advantages for molecular weight determination. Firstly, GFC does not require any chemical labeling or complex sample pretreatment, minimizing interference with sensitive biomolecules. Secondly, GFC can analyze a wide range of molecular weights, accurately handling both small and large molecules, with a relatively simple separation process that does not require extreme conditions. Additionally, GFC provides high resolution, effectively distinguishing components with different molecular weights, making it especially suitable for analyzing complex multi-component samples. Compared to other methods, gel filtration chromatography is gentler, reducing the potential for sample damage, and is particularly ideal for the analysis of biomacromolecules and their complexes. This method has significant application value in protein drug development, vaccine production, and large molecule drug quality control, helping ensure product stability and purity.

 

 

Figure 1. The Principle of Gel Filtration Chromatography

 

Service at MtoZ Biolabs

MtoZ Biolabs offers an efficient Gel Filtration Chromatography Molecular Weight Determination Service, utilizing advanced equipment and technologies to accurately measure the molecular weight of macromolecules such as proteins, nucleic acids, peptides, and other biomolecules. GFC separates different components of a sample by utilizing the distribution of molecules across fillers with varying pore sizes, providing users with clear molecular weight distribution data. This service is applicable to a wide range of biological samples and can analyze the molecular size and aggregation state of complex samples in solution. MtoZ Biolabs integrates GFC with HPLC or other advanced detection technologies to ensure the precision and reliability of molecular weight data, making it widely used in protein purification, characterization, and quality control in various research fields.

 

Service Advantages

1. High Efficiency and Accuracy: Using advanced gel filtration chromatography technology, the service provides fast and accurate molecular weight determination, ensuring high precision and reliability of the data.

 

2. Wide Applicability: This service is suitable for various biomolecules, including proteins, nucleic acids, peptides, and more, handling complex samples to meet diverse research needs.

 

3. Molecular Weight Distribution Analysis: In addition to providing precise molecular weight information, the service offers clear analysis of molecular weight distribution and aggregation state, helping to better understand the structural characteristics of the sample.

 

4. Integration with High-Performance Liquid Chromatography: By combining HPLC with molecular sieving technology, the service enhances separation efficiency and ensures effective separation of different components in the sample.

 

5. Professional Technical Support: MtoZ Biolabs provides expert technical support and data analysis to assist clients in interpreting results and optimizing experimental design.

 

Sample Submission Suggestions

1. Sample Concentration: The sample should have an appropriate concentration to ensure accurate detection during gel filtration. A concentration that is too low may result in reduced resolution, while a concentration that is too high may cause sample aggregation or insufficient separation.

 

2. Sample Purity: The sample should be as pure as possible. Impurities such as solvents, salts, and low molecular weight substances may affect the accuracy of the results. Therefore, preliminary sample purification is recommended to remove solvents and low molecular weight impurities.

 

3. Buffer Solution: The sample should be dissolved in an appropriate buffer. The pH of the buffer should be maintained within the optimal stability range for the protein, and excessive salts or other substances that could interfere with separation should be avoided.

 

4. Particulate Matter: If the sample contains particles or precipitates, they should be removed by centrifugation or filtration to prevent clogging of the chromatography column or interference with the separation process.

 

5. Temperature Requirements: Depending on the nature of the sample, some proteins may need to be stored at low temperatures to prevent degradation or denaturation, especially for heat-sensitive proteins.

 

Case Study

1. Determination of Molecular Size by Size-Exclusion Chromatography (Gel Filtration)

 



Irvine, GB. Curr Protoc Cell Biol. 2001.

 

FAQ

1. How can impurities in the sample (such as salts and low molecular weight substances) be removed to improve the accuracy of the analysis? 

To improve the accuracy of molecular weight determination by gel filtration, impurities can be removed using dialysis or gel filtration columns. Dialysis is effective in removing small molecule impurities (such as salts and solvents) without affecting the structure or function of large molecules like proteins. Another method is to use centrifugal filters or gel filtration columns (e.g., Sephadex) to separate small molecules based on their molecular weight. These methods can remove impurities without damaging the sample structure, ensuring accurate molecular weight determination.

 

2. If the sample concentration is too high, it may lead to molecular aggregation, affecting the chromatographic separation; how can this issue be avoided?

If the sample concentration is too high, it can lead to protein aggregation or precipitation, which can affect separation efficiency. To avoid this, the sample concentration should be appropriately diluted to ensure proper separation during gel filtration. Typically, a concentration range of 0.5–5 mg/mL is recommended to prevent aggregation while maintaining good separation. Additionally, using an appropriate buffer with the correct pH and salt concentration can further help maintain protein stability and prevent aggregation.

 

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Relevant Liquid Chromatography and Mass Spectrometry Parameters

4. The Detailed Information of Molecular Weight Determination 

5. Mass Spectrometry Image

6. Raw Data